| Objective: This study was carried out 1.to research operative skills and experience of rat orthotopic liver transplantation experimental model, to build stable and longtime survival experimental model of rat orthotopic liver transplantation for the next research about acute rejection. 2.to detect morphological change,liver function and relationship between augmenter of liver regeneration (ALR) and INF-γ,IL-2 expression in both isograft and allograft liver transplantation, And then to investigate correlation between ALR and acute rejection.Methods: 1.Modified Kamada two-cuff technique was performed to set up rat orthotopic liver transplantation model, in which suprahepatic vena cava (SHVC) was anastomosed with continuous suture, portal vein (PV) and infrahepatic vena cava (IHVC) were anastomosed with cuff method, arteria hepatica was ligated without reconstruction, and the bile duct was reconstructed by interior stent. Survival rate and general condition of recipients after liver transplantation were observed and operative technique and experience were summarized to direct further experiment. 2.Allogenic (A group) and syngenic (B group) liver transplantation models from Lewis (LEW, Rt11) to Brown-norway (BN, Rt1n) and BN to BN rats were established according to aforesaid method. All survival rats were sacrificed at days 1, 3, 5 and 7 posttransplantation. The pathohistological change of grafts was observed by light microscope and the serum ALT and AST were detected by biochemical analysis. The intra-graft expression of ALR and INF-γ, IL-2 protein and mRNA were evaluated by Real-time PCR and immunohistochemistry staining.Results: 1. Stable and long-time survival Rats orthotopic liver transplantation model was effective constructed with modified Kamada two-cuff technique, and 7 days survival rate was more than 90%. Pathological change of acute rejection was observed at days 3 after liver transplantation from LEW to BN, and typical acute rejection appeared at days 5,7. Similar acute rejection appearance was not observed at same time points from BN to BN. 2. The insult and liver function from both groups was approximate at day 1 after liver transplantation. ALR (1.13±0.10 v.s 1.09±0.12) and IL-2 (0.315±0.088 v.s 0.279±0.048) have no statistical difference, P>0.05. But the expression of INF-γ(0.052±0.009) in allograft group was more significant than that in isograft group (0.007±0.003), P<0.05. The appearance of liver tissue damage and inflammatory cells infiltration at days 3, 5, 7 posttransplantation, as well as the expression of INF-γ(0.477±0.117, 0.162±0.021, 0.122±0.014) and IL-2 (2.718±0.479, 3.330±0.562, 3.506±0.367) mRNA in allogeneic recipients were much more prominent than that in syngeneic recipients, P<0.05. But the expression of ALR (0.59±0.10, 1.57±0.27, 1.98±0.13) in allograft group was decreased compraed to isograft group (0.81±0.11, 2.86±0.37, 3.19±0.35), P<0.05. As the expression of ALR in allogeneic recipients was stepwise promoted, the level of INF-γwas significantly attenuated and IL-2 was gentlely raised instantaneously. Similar change was not detected in graft of syngeneic recipients. 3. Those data revealed obviously negative correlation between ALR and INF-γmRNA levels (r=-0.86, P<0.05) and without correlation between ALR and IL-2 mRNA levels (r=0.322, P>0.05).Conclusion: 1. Modify Kamada two-cuff technique can simplified operative steps and long-time survival rate was efficient improved. 2. Typical acute rejection model was constructed from LEW to BN. 3.ALR may suppress immunocompetent cells activation play an important role in protection on recipients.
|
PDF Full Text Request